RESPONSE OF FROG STOMACH AND SMALL INTESTINE SMOOTH MUSCLE TISSUE TO

ACETYLCHOLINE AND ADRENALINE

Carandang, Lance | Cruz, Reniel | Pasumbal, Romel Jr. | Salem, Andrei | Tolentino, Joshiah
Institute of Biology, University of the Philippines Diliman, Diliman, Quezon City

ABSTRACT

The digestive system which is part of the visceral system of organisms with a vertebra is known to be
innervated by the parasympathetic arm of the nervous system. Smooth muscles in particular responds to
both cholinergic and adrenergic neurotransmitter by increasing and decreasing its contraction per minute
respectively. Both acetylcholine and adrenaline was introduced to the stomach and intestine and it was
compared to the intact organs for reference. Results show that the cholinergic acetylcholine generally
increased the contraction per minute to both the stomach and intestine while adrenaline on the other hand
decreased the contraction per minute of both organs.

INTRODUCTION can cause cause depolarization in adjacent cells,

but neuronal input can manipulate the rate and
The smooth muscle is a non-striated type of strength of contraction. On the other hand,
muscle the forms the wall of hollow organs of multi-unit muscle cells contract only from
the digestive tract and the blood vessels. The neuronal stimulation and each cell acts
lack of striations in the smooth muscle is due to independently (Randall et al., 2002).
the lack of organized groups of actin and myosin
fibers that form sarcomeres found in striated Many of the features of smooth muscles differs
muscle. Myofilaments are gathered in dense from striated muscle. The innervation of smooth
bodies or connected to attachment plaques in the muscle cells is not discrete and intimate; the
sarcolemma. The ɑ-actinin found in the Z disk neurotransmitter is released from varicosities
of a skeletal muscle is also found in high along the axon of the neuron and diffuses over a
concentrations in the attachment plaque, but the distance until it encounters a group of smooth
attachment plaque also contains vinculin which muscle cells with receptor protein distributed
binds to ɑ-actinin and acts as an anchor to the evenly over the cell surface. Smooth muscle
sarcolemma (Randall et al., 2002; Reece et al., cells are generally autonomic and controlled by
2014). hormones, while the single-unit cells are also
sensitive to mechanical stretching that can cause
Smooth muscles can be divided into two depolarization. The response is responsible for
categories: single-unit (unitary) muscles and attaining muscle tension over a length of muscle
multi-unit (multiunitary) muscles. Single-unit cells and peristaltic movement. The contractions
muscles are commonly found in the walls of of smooth muscle is slower and weaker than
visceral organs, and are typically small, spindle- skeletal muscle cells, but can sustain prolonged
shaped, and are connected to each other through periods of contraction (Randall et al., 2002).
electrically conductive gap junctions. Single-
unit muscle cells can contract without stimuli In this experiment, the contractile activity,
from neurons and the depolarization of one cell nervous and chemical control of frog stomach
and small intestine smooth muscles were
1 2 1 2
observed and analyzed.
Intact 3 2 6 5
MATERIALS AND METHODS
Ringer 3 2 5 5
A live frog specimen was procured, and was Acetylcholine 5 - 6 6
prepared for the experiment by double pithing.
The specimen’s abdominal cavity was opened 6 5 8 7
up and smooth muscle contractions of the
stomach and small intestine were observed by Results obtained from the intact visceral systems
counting the number of contractions. of the stomach and intestine show a relative
contraction per minute with the Ringer’s
After observation of the intact muscles, the solution. An increase however, was observed
stomach and portions of the duodenal segment when acetylcholine was introduced to both the
of the small intestine were removed and stomach and intestine. An increase of about 2
submerged under room temperature Ringer’s beats per minute was observed in the stomach
solution. Contractile activity of the smooth while for the intestine, there was increase of
muscles were observed under these conditions about 1 beat per minute.
for one minute.
Table 2. Results of the contraction of the
Afterwards, the smooth muscle samples were stomach and intestine upon the dropwise
exposed to one drop of an acetylcholine solution application of the adrenergic reagent, Adrenaline
and the number of muscle contractions were
counted. Additional drops of acetylcholine were Stomach Intestine
added and subsequent muscle contraction Trial Trial Trial Trial
observation was performed at one minute 1 2 1 2
intervals until a marked difference from activity
under Ringer’s solution was observed. Ringer 2 3 6 5

Adrenaline 2 - 7 4
The samples were resubmerged into room
temperature Ringer’s solution and the procedure 1 0 4 3
used for acetylcholine solutions was repeated
with adrenaline to observe its effect on In contrast to the general increase in the trend of
contractile activity. contraction per minute, application of the
adrenergic reagent decreases the rate of
RESULTS contraction with respect to the ringer solution.

Table 1. Results of the contraction of the

stomach and intestine upon the dropwise
application of the cholinergic reagent,
Acetylcholine DISCUSSION
Stomach Intestine
Smooth muscles lack striations typically seen on
Trial Trial Trial Trial skeletal muscles. Smooth muscles have actin and
myosin fiber, but in a less organized manner stimulation only changes the rhythm and force
where the filaments are gathered in dense bodies of contraction. Hormones, stretching, and local
or connected to attachment plaques in the tissue conditions such as temperature and pH
sarcolemma (Randall et al., 2002). Smooth also affect to contraction of unitary smooth
muscle cells are long fusiform cells that muscle cells (Khurana, 2005).
aggregates into bundles or fasciculi, and
generally contracts involuntarily (Khurana, Multiunit smooth muscle cells, on the other
2005). Like striated muscle, smooth muscle also hand, have individual units that are not
contains a high concentration of ɑ-actinin interconnected by gap junction. Multiunit
attached to the attachment plaques. Vinculin is a smooth muscle cells are located in most blood
unique protein found in smooth muscle cells vessels, epididymis, vas deferens, iris, ciliary
which acts as an anchor to the sarcolemma body, and piloerector muscles. Each cells are
(Randall et al., 2002). innervated by a separate nerve ending, thus the
contraction is neurogenic. Multiunit cells do not
Unlike cardiac and skeletal muscle, smooth contract autonomously and relies on stimuli
muscle does not contain the calcium-binding from nerves (Khurana, 2005).
protein troponin. Contraction is initiated by a
calcium-regulated phosphorylation of myosin, Smooth muscle cells are innervated by both
rather than a calcium-activated troponin system. sympathetic and parasympathetic nerve. As the
Instead, elevated intracellular Ca2+ levels end of the nerves approach the smooth muscle
regulate contraction by a mechanism that cells, extensive branches with enlargements
depends on calmodulin (Alberts, 2015). Smooth called varicosities innervate each cell. The
muscles lack sarcomeres but actin filaments are varicosities release neurotransmitters
arranged obliquely in the cytoplasm. Striations (acetylcholine or adrenaline) that diffuse into the
would not also be present because of the lack of interstitial fluid and into the cells (Khurana,
sarcomeres. But despite the lack of sarcomeres, 2005).
contraction in smooth muscle cells is still
possible. Contraction in smooth muscle cells are Acetylcholine, in response to the
achieved via the shortening of intermediate parasympathetic nerves, mediates contraction of
filaments that link two dense bodies (Webb, smooth muscle cells (Randall et al., 2002). On
2003). It is also triggered by an influx of the other hand, adrenaline will respond
calcium ions (Alberts, 2015). differently to smooth muscles depending on the
location of the organ and the receptors. For
Smooth muscles are of two types: single-unit example, adrenaline stimulates smooth muscle
(unitary) and multiunit (multiunitary) smooth contraction in the ureter, vas deferens, uterus,
muscles. Single-unit cells are present in the urethral sphincter, urothelium, bronchioles and
walls of hollow viscera such as the arrector pili muscle in hairs which are affected
gastrointestinal tract, uterus, urinary bladder, by α1-adrenergic receptors (Schmitz et al.,
and respiratory tract. Smooth muscle cells are 1981). For the stomach, it is affected by β 1
joined by low resistance gap junction with high receptor which increases ghrelin secretion from
conductance for ions. The syncytial nature of the stomach (Zhao et al., 2010).
unitary muscle cells allows a large area to
contract as a single unit. The contraction of In the cardiac muscle, acetylcholine increases
unitary cells are myogenic and nervous potassium conductance of the pacemaker cells
(Randall et al., 2002). Acetylcholine causes as well in order to boost the cardiac output.
bradycardia, and decrease of the strength of parasympathetic to viscera of thorax and
atrial contractions. The cellular abdomen (Sherwood, 2013). Excessive vagus
hyperpolarization resulting from opening of nerve stimulation will lead to bradycardia and
potassium channels linked to the G proteins and AV node blocks (Marieb, 2009).
potassium exit out of the cell, inducing an
increase in polarization, causes cardiac muscle On the other hand, the vagus nerve has a main
inhibition. The decrease of the force of role in three phases of gastric secretion. During
contraction comes from a decrease of Ca 2+ entry cephalic phase, parasympathetic action
in the cell, probably by inhibition of potentials are carried by the vagus nerves to the
adenylcyclase. Hence, intravenous injection of stomach. The preganglionic parasympathetic
very low doses of acetylcholine in animals or in vagus nerve fibers stimulate postganglionic
human beings causes immediate and fugacious neurons in the enteric plexus of the stomach
fall of the arterial pressure resulting from while the postganglionic parasympathetic vagus
cardiac slowing and vasodilation (Chamales et nerve fibers will stimulate secretions by parietal
al., 1975). On the other hand, adrenaline reacts and chief cells and stimulate gastrin secretions.
with both α- and β-adrenoreceptors, causing During gastric phase, the enlargement of the
vasoconstriction and vasodilation, respectively. stomach activates a parasympathetic reflex. The
In the skeletal muscle, acetylcholine is able to action potentials are carried by the vagus nerves
bind to acetylcholine receptors and open ligand to the medulla oblongata. Lastly, afferent vagal
gated sodium channels allowing for sodium ions action potentials inhibit efferent action potentials
to enter the cell and stimulate contraction. from the medulla oblongata which causes
Adrenaline is able to stimulate β-adrenotropic inhibition of gastric secretions in the stomach
receptors inducing glucose uptake but has (Marieb, 2009).
different effects on different muscle types. It has
been found that adrenaline is able to increase CONCLUSION
contraction in fast-twitch muscles and has an
opposite effect in slow-twitch muscles (Öberg, Based from the results obtained from the
2013). experiment, it can be concluded that there is a
general increase in the contraction per minute of
The vagus nerve, a major nerve of the the organs composed of smooth muscle tissues.
parasympathetic nervous system, innervates Acetylcholine, which is a known cholinergic
both the gastrointestinal tract and the heart reagent, stimulated the parasympathetic arm of
which cause different responses for smooth and the nerves triggering stimulation that resulted in
cardiac muscles, respectively. The vagus nerve the increase in the contraction per minute.
is the tenth cranial nerve that innervates organs Alternatively, adrenaline which is a potent
in the thoracic and abdominal cavities adrenergic reagent decrease the contraction per
(Sherwood, 2013). The vagus nerve helps minute of both intestine and stomach due to the
regulate the functions of the thoracic and fact that it stimulated the sympathetic arm of the
abdominal organs such as heart rate, respiration visceral systems.
rate, and digestion. (Marieb, 2009). The vagus
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